DOCSLIB.ORG

JNK Activation of BIM Promotes Hepatic Oxidative Stress, Steatosis and Insulin Resistance in Obesity

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
JNK Activation of BIM Promotes Hepatic Oxidative Stress, Steatosis and Insulin Resistance in Obesity Page 1 of 62 Diabetes JNK activation of BIM promotes hepatic oxidative stress, steatosis and insulin resistance in obesity. Sara A. Litwak1, Lokman Pang1,2, Sandra Galic1, Mariana Igoillo-Esteve3, William J. Stanley1,2, Jean-Valery Turatsinze3, Kim Loh1, Helen E. Thomas1,2, Arpeeta Sharma4, Eric Trepo5,6, Christophe Moreno5,6, Daniel J. Gough7,8, Decio L. Eizirik3, Judy B. de Haan4, Esteban N. Gurzov1,2,a 1St Vincent’s Institute of Medical Research, Melbourne, Australia. 2Department of Medicine, St. Vincent’s Hospital, The University of Melbourne, Melbourne, Australia. 3ULB Center for Diabetes Research, Université Libre de Bruxelles (ULB), Brussels, Belgium. 4Oxidative Stress Laboratory, Basic Science Division, Baker Heart and Diabetes Institute, Melbourne, VIC, Australia. 5CUB Hôpital Erasme, Université Libre de Bruxelles (ULB), Belgium. 6Laboratory of experimental Gastroenterology, Université Libre de Bruxelles (ULB), Belgium. 7Hudson Institute of Medical Research, Clayton, VIC, Australia. 8Department of Molecular and Translational Science, Monash University, Clayton, Vic, Australia. aPresent address, to where correspondence and reprint requests should be addressed: Dr Esteban N. Gurzov ULB Center for Diabetes Research Université Libre de Bruxelles Campus Erasme, Route de Lennik 808, B-1070-Brussels-Belgium Phone: +32 2 5556242 Fax: +32 2 5556239 Email: [email protected] Disclosure statement: The authors declare no conflict of interest Running Title: BIM regulates lipid metabolism in hepatocytes. 1 Diabetes Publish Ahead of Print, published online September 19, 2017 Diabetes Page 2 of 62 ABSTRACT The BCL-2 family are crucial regulators of the mitochondrial pathway of apoptosis in normal physiology and disease. Besides their role in cell death, BCL-2 proteins have been implicated in the regulation of mitochondrial oxidative phosphorylation and cellular metabolism. It remains unclear, however, whether these proteins have a physiological role in glucose homeostasis and metabolism in vivo. Here we report that fat accumulation in the liver increases JNK-dependent BIM expression in hepatocytes. To determine the consequences of hepatic BIM deficiency in diet- induced obesity, we generated liver-specific BIM knockout (BLKO) mice. BLKO mice had lower hepatic lipid content, increased insulin signalling and improved global glucose metabolism. Consistent with these findings, lipogenic and lipid uptake genes were downregulated and lipid oxidation enhanced in obese BLKO mice. Mechanistically, BIM deficiency improved mitochondrial function and decreased oxidative stress, oxidation of protein tyrosine phosphatases and ameliorated activation of PPAR-γ/SREBP1/CD36 in hepatocytes from high fat fed mice. Importantly, short- term knockdown of BIM rescued obese mice from insulin resistance, evidenced by reduced fat accumulation and improved insulin sensitivity. Our data indicate that BIM is an important regulator of liver dysfunction in obesity, and a novel therapeutic target for restoring hepatocyte function. 2 Page 3 of 62 Diabetes INTRODUCTION Obesity is a major risk factor for the development of severe complications such as non- alcoholic fatty liver disease (NAFLD), cardiovascular disease and diabetes (1). The liver has an essential role in the regulation of glucose homeostasis, producing glucose during fasting periods to prevent hypoglycaemia, maintaining brain function and survival. In obese individuals, excessive nutrients are stored in the liver as fat droplets (hepatosteatosis), which can eventually result in inflammation, insulin resistance and diabetes (2; 3). Obesity triggers activation of the c-Jun N-terminal kinase (JNK) pathway in hepatocytes with subsequent insulin resistance as well as steatosis (4; 5). The mechanisms of obesity-induced JNK activation include endoplasmic reticulum (ER) and oxidative stress, Toll-like receptors (TLRs) and inflammatory cytokines, such as tumor necrosis factor (TNF)-α (6). Hepatocyte‐specific deletion of both JNK1 and JNK2 improved glucose and insulin tolerance, increased hepatic insulin action and lowered fasting blood glucose levels in obese mice (7). However, the molecular pathways and proteins mediating JNK-induced hepatocyte dysfunction remain to be fully elucidated. The BCL-2 proteins are key regulators of apoptotic pathways (8), but growing evidence indicates that these proteins can also play an important role in glucose homeostasis and metabolism (9; 10). We have recently reported that loss of the BCL- 2 protein PUMA influences circulating leptin levels and food intake in mice (11). Moreover, BCL-2-associated death promoter (BAD) phosphorylation activates glucokinase and thus controls hepatic gluconeogenesis and insulin secretion in pancreatic β-cells (12; 13). In addition, BCL-2 proteins have been reported to stimulate glucose consumption (14) and regulate glucose metabolism through 3 Diabetes Page 4 of 62 mitochondrial activity (15) or Ca2+ trafficking (16; 17). This dual role in metabolism/apoptosis of BCL-2 proteins is reminiscent of the action of cytochrome c, a protein that functions as a crucial component for oxidative phosphorylation in the mitochondria, but when displaced to the cytosol induces cleavage of caspases and cell death (9). BCL-2 interacting mediator of cell death (BIM) is a BCL-2 homology 3 (BH3)-only protein widely expressed in tissues (18). Saturated free fatty acids trigger cell death in hepatocytes in culture through BIM up-regulation (19-21). Fat accumulation and hepatocellular carcinoma in liver-specific STAT5 knockout mice is associated with down-regulation of BIM and PUMA (22). In line with these findings, liver apoptosis in BCL-XL- or MCL-1-knockout mice is ameliorated by simultaneous BIM deletion (23). While it is well accepted that BIM has a role in apoptosis induction in hepatocytes, its role in the regulation of hepatic lipid and glucose metabolism in vivo is unclear. Moreover, it remains unknown whether dysregulation of BIM may contribute to the development of metabolic diseases such as liver steatosis and insulin resistance. Here, we show that BIM is induced in hepatocytes during obesity. To clarify the role of BIM, we generated a novel liver-specific BIM-deficient mouse model and observed that BIM deletion decreases fat accumulation, and improves insulin signalling in vivo. These novel observations highlight BIM as a key mediator of liver dysfunction in obesity. RESEARCH DESIGN AND METHODS Human samples. We studied 19 biopsy specimens of patients undergoing a liver biopsy for medical 4 Page 5 of 62 Diabetes reasons in our institution. The clinical characteristics of these patients are shown in Supplementary Figure 1E. The biopsies were collected after approval of the Erasme Hospital Ethics Committee. Written informed consent was obtained from each participant. Mice. Mice were maintained at St. Vincent’s Institute animal care facility on a 12h-light- dark cycle in a temperature-controlled room and obtained food and water ad libitum. Bimlox/lox mice were generated on a C57BL/6 background as previously described (24). Tissue-specific deletion of BIM was generated by crossing Bimlox/lox mice with Alb-Cre (C57BL/6) mice (JAX). Male mice were kept on regular chow (20% protein, 6% fat and 3.2% crude fibre) or placed at 8-10 weeks of age on a high fat diet (SF04- 027 Speciality Feeds, Perth, Western Australia) for 14-20 weeks. The nutritional composition of the high fat diet was 18.4% protein, 23.5% fat and 4.7% crude fibre. The calculated composition of fatty acids in the high fat diet is: 14.31% total saturated fats, 7.54% total mono-unsaturated fats, and 2.07% total polyunsaturated fats. In this diet, 46% of total energy is from lipids, 20% of total calculated energy from protein and the remainder from carbohydrates. All animal studies were conducted at St Vincent’s Institute (Melbourne, Australia) following the guidelines of the Institutional Animal Ethics Committee. Metabolic analysis. After 14-15 weeks on the diet and 24h of acclimatisation, oxygen consumption, energy expenditure, respiratory exchange ratio, activity and food intake were assessed using a Comprehensive Lab Animal Monitoring System (CLAMS, Columbus- 5 Diabetes Page 6 of 62 Instruments, OH). Data was averaged for 2 dark and light cycles. Body and tissue composition were determined by MRI scanning (EchoMRI, Houston, TX). After 18- 20 weeks of chow/high fat feeding, mice were sacrificed and tissues were weighed, and collected for further analysis. Serum triacylglycerides (Wako Diagnostic, Richmond, VA), insulin (Merck Millipore, Billerica, MA), and β-hydroxybutyrate (Sigma-Aldrich, Darmstadt, Germany) concentrations were determined using commercial kits and following manufacturer’s instructions. Mice were fasted for 4h before performing intra-peritoneal insulin tolerance test. Insulin (Actrapid, Novo Nordisk, Bagsværd, Denmark), at a dose of 0.65mU/g, was injected intraperitoneally and blood glucose was measured after tail bleeding. Mice were fasted for 6h before an intra-peritoneal glucose (2mg/g, Baxter, Deerfield, IL) or pyruvate (2mg/g, Sigma-Aldrich) tolerance test. Cell culture and treatments. Mouse hepatocytes were isolated by a two-step collagenase A (0.05% w/v; Roche Diagnostics) perfusion method as described previously (25). Where indicated cells were treated with 0.5mM sodium palmitate in the presence of 1% w/v fatty acid free BSA (26). The JNK inhibitor SP600125 (Santa Cruz Biotechnology, CA) was used at 10µM (27). JNKs (JNK1 and JNK2) were knocked down transiently
Load more

Recommended publications
  • Genome-Wide Association Study of Behavioral Disinhibition in a Selected Adolescent Sample Jaime Derringer *, Robin P. Corley
    Genome-wide association study of behavioral disinhibition in a selected adolescent sample Jaime Derringer1*, Robin P. Corley2, Brett C. Haberstick2, Susan E. Young2, Brittany Demmitt2, Daniel P. Howrigan3, Robert M. Kirkpatrick4, William G. Iacono5, Matt McGue5, Matthew Keller2, Sandra Brown6, Susan Tapert6, Christian J. Hopfer7, Michael C. Stallings2, Thomas J. Crowley7, Soo Hyun Rhee2, Ken Krauter2, John K. Hewitt2, Matthew B. McQueen2 1 University of Illinois Urbana-Champaign, 2 University of Colorado Boulder, 3 Massachusetts General Hospital/Harvard Medical School, 4 Virginia Commonwealth University, 5 University of Minnesota, 6 University of California San Diego, 7 University of Colorado Denver * Corresponding Author: Jaime Derringer, [email protected], Department of Psychology, University of Illinois Urbana-Champaign, Champaign IL USA 61820 Behavioral disinhibition GWAS 2 Abstract Behavioral disinhibition (BD) is a quantitative measure designed to capture the heritable variation encompassing risky and impulsive behaviors. As a result, BD represents an ideal target for discovering genetic loci that predispose individuals to a wide range of antisocial behaviors and substance misuse that together represent a large cost to society as a whole. Published genome-wide association studies (GWAS) have examined specific phenotypes that fall under the umbrella of BD (e.g. alcohol dependence, conduct disorder); however no GWAS has specifically examined the overall BD construct. We conducted a GWAS of BD using a sample of 1,901 adolescents over-selected for characteristics that define high BD, such as substance and antisocial behavior problems, finding no individual locus that surpassed genome-wide significance. Although no single SNP was significantly associated with BD, restricted maximum likelihood analysis estimated that 49.3% of the variance in BD within the Caucasian sub-sample was accounted for by the genotyped SNPs (p=0.06).
    [Show full text]
  • Global-Scale Analysis of the Dynamic Transcriptional Adaptations Within Skeletal Muscle During Hypertrophic Growth
    University of Kentucky UKnowledge Theses and Dissertations--Physiology Physiology 2015 GLOBAL-SCALE ANALYSIS OF THE DYNAMIC TRANSCRIPTIONAL ADAPTATIONS WITHIN SKELETAL MUSCLE DURING HYPERTROPHIC GROWTH Tyler Kirby University of Kentucky, [email protected] Right click to open a feedback form in a new tab to let us know how this document benefits ou.y Recommended Citation Kirby, Tyler, "GLOBAL-SCALE ANALYSIS OF THE DYNAMIC TRANSCRIPTIONAL ADAPTATIONS WITHIN SKELETAL MUSCLE DURING HYPERTROPHIC GROWTH" (2015). Theses and Dissertations--Physiology. 22. https://uknowledge.uky.edu/physiology_etds/22 This Doctoral Dissertation is brought to you for free and open access by the Physiology at UKnowledge. It has been accepted for inclusion in Theses and Dissertations--Physiology by an authorized administrator of UKnowledge. For more information, please contact [email protected]. STUDENT AGREEMENT: I represent that my thesis or dissertation and abstract are my original work. Proper attribution has been given to all outside sources. I understand that I am solely responsible for obtaining any needed copyright permissions. I have obtained needed written permission statement(s) from the owner(s) of each third-party copyrighted matter to be included in my work, allowing electronic distribution (if such use is not permitted by the fair use doctrine) which will be submitted to UKnowledge as Additional File. I hereby grant to The University of Kentucky and its agents the irrevocable, non-exclusive, and royalty-free license to archive and make accessible my work in whole or in part in all forms of media, now or hereafter known. I agree that the document mentioned above may be made available immediately for worldwide access unless an embargo applies.
    [Show full text]
  • Role of Mitochondrial Ribosomal Protein S18-2 in Cancerogenesis and in Regulation of Stemness and Differentiation
    From THE DEPARTMENT OF MICROBIOLOGY TUMOR AND CELL BIOLOGY (MTC) Karolinska Institutet, Stockholm, Sweden ROLE OF MITOCHONDRIAL RIBOSOMAL PROTEIN S18-2 IN CANCEROGENESIS AND IN REGULATION OF STEMNESS AND DIFFERENTIATION Muhammad Mushtaq Stockholm 2017 All previously published papers were reproduced with permission from the publisher. Published by Karolinska Institutet. Printed by E-Print AB 2017 © Muhammad Mushtaq, 2017 ISBN 978-91-7676-697-2 Role of Mitochondrial Ribosomal Protein S18-2 in Cancerogenesis and in Regulation of Stemness and Differentiation THESIS FOR DOCTORAL DEGREE (Ph.D.) By Muhammad Mushtaq Principal Supervisor: Faculty Opponent: Associate Professor Elena Kashuba Professor Pramod Kumar Srivastava Karolinska Institutet University of Connecticut Department of Microbiology Tumor and Cell Center for Immunotherapy of Cancer and Biology (MTC) Infectious Diseases Co-supervisor(s): Examination Board: Professor Sonia Lain Professor Ola Söderberg Karolinska Institutet Uppsala University Department of Microbiology Tumor and Cell Department of Immunology, Genetics and Biology (MTC) Pathology (IGP) Professor George Klein Professor Boris Zhivotovsky Karolinska Institutet Karolinska Institutet Department of Microbiology Tumor and Cell Institute of Environmental Medicine (IMM) Biology (MTC) Professor Lars-Gunnar Larsson Karolinska Institutet Department of Microbiology Tumor and Cell Biology (MTC) Dedicated to my parents ABSTRACT Mitochondria carry their own ribosomes (mitoribosomes) for the translation of mRNA encoded by mitochondrial DNA. The architecture of mitoribosomes is mainly composed of mitochondrial ribosomal proteins (MRPs), which are encoded by nuclear genomic DNA. Emerging experimental evidences reveal that several MRPs are multifunctional and they exhibit important extra-mitochondrial functions, such as involvement in apoptosis, protein biosynthesis and signal transduction. Dysregulations of the MRPs are associated with severe pathological conditions, including cancer.
    [Show full text]
  • A Computational Approach for Defining a Signature of Β-Cell Golgi Stress in Diabetes Mellitus
    Page 1 of 781 Diabetes A Computational Approach for Defining a Signature of β-Cell Golgi Stress in Diabetes Mellitus Robert N. Bone1,6,7, Olufunmilola Oyebamiji2, Sayali Talware2, Sharmila Selvaraj2, Preethi Krishnan3,6, Farooq Syed1,6,7, Huanmei Wu2, Carmella Evans-Molina 1,3,4,5,6,7,8* Departments of 1Pediatrics, 3Medicine, 4Anatomy, Cell Biology & Physiology, 5Biochemistry & Molecular Biology, the 6Center for Diabetes & Metabolic Diseases, and the 7Herman B. Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, IN 46202; 2Department of BioHealth Informatics, Indiana University-Purdue University Indianapolis, Indianapolis, IN, 46202; 8Roudebush VA Medical Center, Indianapolis, IN 46202. *Corresponding Author(s): Carmella Evans-Molina, MD, PhD ([email protected]) Indiana University School of Medicine, 635 Barnhill Drive, MS 2031A, Indianapolis, IN 46202, Telephone: (317) 274-4145, Fax (317) 274-4107 Running Title: Golgi Stress Response in Diabetes Word Count: 4358 Number of Figures: 6 Keywords: Golgi apparatus stress, Islets, β cell, Type 1 diabetes, Type 2 diabetes 1 Diabetes Publish Ahead of Print, published online August 20, 2020 Diabetes Page 2 of 781 ABSTRACT The Golgi apparatus (GA) is an important site of insulin processing and granule maturation, but whether GA organelle dysfunction and GA stress are present in the diabetic β-cell has not been tested. We utilized an informatics-based approach to develop a transcriptional signature of β-cell GA stress using existing RNA sequencing and microarray datasets generated using human islets from donors with diabetes and islets where type 1(T1D) and type 2 diabetes (T2D) had been modeled ex vivo. To narrow our results to GA-specific genes, we applied a filter set of 1,030 genes accepted as GA associated.
    [Show full text]
  • CSNK2B Monoclonal Antibody Catalog Number:67866-1-Ig
    For Research Use Only CSNK2B Monoclonal antibody www.ptgcn.com Catalog Number:67866-1-Ig Catalog Number: GenBank Accession Number: CloneNo.: Basic Information 67866-1-Ig BC112017 1B5A6 Size: GeneID (NCBI): Recommended Dilutions: 1000 μg/ml 1460 WB 1:5000-1:20000 Source: Full Name: IF 1:200-1:800 Mouse casein kinase 2, beta polypeptide Isotype: Calculated MW: IgG1 215 aa, 25 kDa Purification Method: Observed MW: Protein G purification 27 kDa Immunogen Catalog Number: AG19180 Applications Tested Applications: Positive Controls: IF, WB,ELISA WB : A549 cells; LNCaP cells, HeLa cells, Jurkat cells, Species Specificity: pig brain tissue, rat brain tissue, mouse brain tissue Human, mouse, rat, pig IF : HeLa cells; CSNK2B is a ubiquitous protein kinase which regulates metabolic pathways, signal transduction, transcription, Background Information translation, and replication. The enzyme is composed of three subunits, alpha, alpha prime and beta, which form a tetrameric holoenzyme. The alpha and alpha prime subunits are catalytic, while the beta subunit serves regulatory functions. The enzyme localizes to the endoplasmic reticulum and the Golgi apparatus. It participates in Wnt signaling, and plays a complex role in regulating the basal catalytic activity of the alpha subunit. Storage: Storage Store at -20ºC. Stable for one year after shipment. Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3. Aliquoting is unnecessary for -20ºC storage For technical support and original validation data for this product please contact: This product is exclusively available under Proteintech T: 4006900926 E: [email protected] W: ptgcn.com Group brand and is not available to purchase from any other manufacturer.
    [Show full text]
  • Integrating Single-Step GWAS and Bipartite Networks Reconstruction Provides Novel Insights Into Yearling Weight and Carcass Traits in Hanwoo Beef Cattle
    animals Article Integrating Single-Step GWAS and Bipartite Networks Reconstruction Provides Novel Insights into Yearling Weight and Carcass Traits in Hanwoo Beef Cattle Masoumeh Naserkheil 1 , Abolfazl Bahrami 1 , Deukhwan Lee 2,* and Hossein Mehrban 3 1 Department of Animal Science, University College of Agriculture and Natural Resources, University of Tehran, Karaj 77871-31587, Iran; [email protected] (M.N.); [email protected] (A.B.) 2 Department of Animal Life and Environment Sciences, Hankyong National University, Jungang-ro 327, Anseong-si, Gyeonggi-do 17579, Korea 3 Department of Animal Science, Shahrekord University, Shahrekord 88186-34141, Iran; [email protected] * Correspondence: [email protected]; Tel.: +82-31-670-5091 Received: 25 August 2020; Accepted: 6 October 2020; Published: 9 October 2020 Simple Summary: Hanwoo is an indigenous cattle breed in Korea and popular for meat production owing to its rapid growth and high-quality meat. Its yearling weight and carcass traits (backfat thickness, carcass weight, eye muscle area, and marbling score) are economically important for the selection of young and proven bulls. In recent decades, the advent of high throughput genotyping technologies has made it possible to perform genome-wide association studies (GWAS) for the detection of genomic regions associated with traits of economic interest in different species. In this study, we conducted a weighted single-step genome-wide association study which combines all genotypes, phenotypes and pedigree data in one step (ssGBLUP). It allows for the use of all SNPs simultaneously along with all phenotypes from genotyped and ungenotyped animals. Our results revealed 33 relevant genomic regions related to the traits of interest.
    [Show full text]
  • DDX47 Polyclonal Antibody
    PRODUCT DATA SHEET Bioworld Technology,Inc. DDX47 polyclonal antibody Catalog: BS71653 Host: Rabbit Reactivity: Human,Mouse,Rat BackGround: munogen and the purity is > 95% (by SDS-PAGE). This gene encodes a member of the DEAD box protein Applications: family. DEAD box proteins, characterized by the con- WB 1:500 - 1:2000 served motif Asp-Glu-Ala-Asp (DEAD), are putative IHC 1:50 - 1:200 RNA helicases. They are implicated in a number of cellu- Storage&Stability: lar processes involving alteration of RNA secondary Store at 4°C short term. Aliquot and store at -20°C long structure, such as translation initiation, nuclear and mito- term. Avoid freeze-thaw cycles. chondrial splicing, and ribosome and spliceosome assem- Specificity: bly. Based on their distribution patterns, some members DDX47 polyclonal antibody detects endogenous levels of of this family are believed to be involved in embryogene- DDX47 protein. sis, spermatogenesis, and cellular growth and division. DATA: The protein encoded by this gene can shuttle between the nucleus and the cytoplasm, and has an RNA-independent ATPase activity. Two alternatively spliced transcript var- iants encoding distinct isoforms have been found for this gene. Product: Rabbit IgG, 1mg/ml in PBS with 0.02% sodium azide, 50% glycerol, pH7.2 Western blot analysis of extracts of various cell lines, using DDX47 an- Molecular Weight: tibody. ~ 50 kDa Note: Swiss-Prot: For research use only, not for use in diagnostic procedure. Q9H0S4 Purification&Purity: The antibody was affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific im- Bioworld Technology, Inc. Bioworld technology, co. Ltd.
    [Show full text]
  • De Novo EIF2AK1 and EIF2AK2 Variants Are Associated with Developmental Delay, Leukoencephalopathy, and Neurologic Decompensation
    bioRxiv preprint doi: https://doi.org/10.1101/757039; this version posted September 16, 2019. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. De novo EIF2AK1 and EIF2AK2 variants are associated with developmental delay, leukoencephalopathy, and neurologic decompensation Dongxue Mao1,2, Chloe M. Reuter3,4, Maura R.Z. Ruzhnikov5,6, Anita E. Beck7, Emily G. Farrow8,9,10, Lisa T. Emrick1,11,12,13, Jill A. Rosenfeld12, Katherine M. Mackenzie5, Laurie Robak2,12,13, Matthew T. Wheeler3,14, Lindsay C. Burrage12,13, Mahim Jain15, Pengfei Liu12, Daniel Calame11,13, Sebastien Küry17,18, Martin Sillesen19, Klaus Schmitz-Abe20, Davide Tonduti21, Luigina Spaccini22, Maria Iascone23, Casie A. Genetti20, Madeline Graf16, Alyssa Tran12, Mercedes Alejandro12, Undiagnosed Diseases Network, Brendan H. Lee12,13, Isabelle Thiffault8,9,24, Pankaj B. Agrawal#,20, Jonathan A. Bernstein#,3,25, Hugo J. Bellen#,2,12,26,27,28, Hsiao- Tuan Chao#,1,2,11,12,13,28,27,29 #Correspondence should be addressed: [email protected] (P.A.), [email protected] (J.A.B.), [email protected] (H.J.B.), [email protected] (H.T.C.) 1Department of Pediatrics, Baylor College of Medicine (BCM), Houston, TX 2Jan and Dan Duncan Neurological Research Institute, Texas Children’s Hospital, Houston, TX 3Stanford Center for Undiagnosed Diseases, Stanford University, Stanford, CA 4Stanford Center for Inherited Cardiovascular Disease, Division of Cardiovascular Medicine,
    [Show full text]
  • Molecular Characterization of Acute Myeloid Leukemia by Next Generation Sequencing: Identification of Novel Biomarkers and Targets of Personalized Therapies
    Alma Mater Studiorum – Università di Bologna Dipartimento di Medicina Specialistica, Diagnostica e Sperimentale Dottorato di Ricerca in Oncologia, Ematologia e Patologia XXX Ciclo Settore Scientifico Disciplinare: MED/15 Settore Concorsuale:06/D3 Molecular characterization of acute myeloid leukemia by Next Generation Sequencing: identification of novel biomarkers and targets of personalized therapies Presentata da: Antonella Padella Coordinatore Prof. Pier-Luigi Lollini Supervisore: Prof. Giovanni Martinelli Esame finale anno 2018 Abstract Acute myeloid leukemia (AML) is a hematopoietic neoplasm that affects myeloid progenitor cells and it is one of the malignancies best studied by next generation sequencing (NGS), showing a highly heterogeneous genetic background. The aim of the study was to characterize the molecular landscape of 2 subgroups of AML patients carrying either chromosomal number alterations (i.e. aneuploidy) or rare fusion genes. We performed whole exome sequencing and we integrated the mutational data with transcriptomic and copy number analysis. We identified the cell cycle, the protein degradation, response to reactive oxygen species, energy metabolism and biosynthetic process as the pathways mostly targeted by alterations in aneuploid AML. Moreover, we identified a 3-gene expression signature including RAD50, PLK1 and CDC20 that characterize this subgroup. Taking advantage of RNA sequencing we aimed at the discovery of novel and rare gene fusions. We detected 9 rare chimeric transcripts, of which partner genes were transcription factors (ZEB2, BCL11B and MAFK) or tumor suppressors (SAV1 and PUF60) rarely translocated across cancer types. Moreover, we detected cryptic events hiding the loss of NF1 and WT1, two recurrently altered genes in AML. Finally, we explored the oncogenic potential of the ZEB2-BCL11B fusion, which revealed no transforming ability in vitro.
    [Show full text]
  • The Role of Cyclin B3 in Mammalian Meiosis
    THE ROLE OF CYCLIN B3 IN MAMMALIAN MEIOSIS by Mehmet Erman Karasu A Dissertation Presented to the Faculty of the Louis V. Gerstner Jr. Graduate School of Biomedical Sciences, Memorial Sloan Kettering Cancer Center In Partial Fulfillment of the Requirements for the Degree of Doctor of Philosophy New York, NY November, 2018 Scott Keeney, PhD Date Dissertation Mentor Copyright © Mehmet Erman Karasu 2018 DEDICATION I would like to dedicate this thesis to my parents, Mukaddes and Mustafa Karasu. I have been so lucky to have their support and unconditional love in this life. ii ABSTRACT Cyclins and cyclin dependent kinases (CDKs) lie at the center of the regulation of the cell cycle. Cyclins as regulatory partners of CDKs control the switch-like cell cycle transitions that orchestrate orderly duplication and segregation of genomes. Similar to somatic cell division, temporal regulation of cyclin-CDK activity is also important in meiosis, which is the specialized cell division that generates gametes for sexual production by halving the genome. Meiosis does so by carrying out one round of DNA replication followed by two successive divisions without another intervening phase of DNA replication. In budding yeast, cyclin-CDK activity has been shown to have a crucial role in meiotic events such as formation of meiotic double-strand breaks that initiate homologous recombination. Mammalian cells express numerous cyclins and CDKs, but how these proteins control meiosis remains poorly understood. Cyclin B3 was previously identified as germ cell specific, and its restricted expression pattern at the beginning of meiosis made it an interesting candidate to regulate meiotic events.
    [Show full text]
  • Mechanisms of Synaptic Plasticity Mediated by Clathrin Adaptor-Protein Complexes 1 and 2 in Mice
    Mechanisms of synaptic plasticity mediated by Clathrin Adaptor-protein complexes 1 and 2 in mice Dissertation for the award of the degree “Doctor rerum naturalium” at the Georg-August-University Göttingen within the doctoral program “Molecular Biology of Cells” of the Georg-August University School of Science (GAUSS) Submitted by Ratnakar Mishra Born in Birpur, Bihar, India Göttingen, Germany 2019 1 Members of the Thesis Committee Prof. Dr. Peter Schu Institute for Cellular Biochemistry, (Supervisor and first referee) University Medical Center Göttingen, Germany Dr. Hans Dieter Schmitt Neurobiology, Max Planck Institute (Second referee) for Biophysical Chemistry, Göttingen, Germany Prof. Dr. med. Thomas A. Bayer Division of Molecular Psychiatry, University Medical Center, Göttingen, Germany Additional Members of the Examination Board Prof. Dr. Silvio O. Rizzoli Department of Neuro-and Sensory Physiology, University Medical Center Göttingen, Germany Dr. Roland Dosch Institute of Developmental Biochemistry, University Medical Center Göttingen, Germany Prof. Dr. med. Martin Oppermann Institute of Cellular and Molecular Immunology, University Medical Center, Göttingen, Germany Date of oral examination: 14th may 2019 2 Table of Contents List of abbreviations ................................................................................. 5 Abstract ................................................................................................... 7 Chapter 1: Introduction ............................................................................
    [Show full text]
  • WO 2019/079361 Al 25 April 2019 (25.04.2019) W 1P O PCT
    (12) INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT) (19) World Intellectual Property Organization I International Bureau (10) International Publication Number (43) International Publication Date WO 2019/079361 Al 25 April 2019 (25.04.2019) W 1P O PCT (51) International Patent Classification: CA, CH, CL, CN, CO, CR, CU, CZ, DE, DJ, DK, DM, DO, C12Q 1/68 (2018.01) A61P 31/18 (2006.01) DZ, EC, EE, EG, ES, FI, GB, GD, GE, GH, GM, GT, HN, C12Q 1/70 (2006.01) HR, HU, ID, IL, IN, IR, IS, JO, JP, KE, KG, KH, KN, KP, KR, KW, KZ, LA, LC, LK, LR, LS, LU, LY, MA, MD, ME, (21) International Application Number: MG, MK, MN, MW, MX, MY, MZ, NA, NG, NI, NO, NZ, PCT/US2018/056167 OM, PA, PE, PG, PH, PL, PT, QA, RO, RS, RU, RW, SA, (22) International Filing Date: SC, SD, SE, SG, SK, SL, SM, ST, SV, SY, TH, TJ, TM, TN, 16 October 2018 (16. 10.2018) TR, TT, TZ, UA, UG, US, UZ, VC, VN, ZA, ZM, ZW. (25) Filing Language: English (84) Designated States (unless otherwise indicated, for every kind of regional protection available): ARIPO (BW, GH, (26) Publication Language: English GM, KE, LR, LS, MW, MZ, NA, RW, SD, SL, ST, SZ, TZ, (30) Priority Data: UG, ZM, ZW), Eurasian (AM, AZ, BY, KG, KZ, RU, TJ, 62/573,025 16 October 2017 (16. 10.2017) US TM), European (AL, AT, BE, BG, CH, CY, CZ, DE, DK, EE, ES, FI, FR, GB, GR, HR, HU, ΓΕ , IS, IT, LT, LU, LV, (71) Applicant: MASSACHUSETTS INSTITUTE OF MC, MK, MT, NL, NO, PL, PT, RO, RS, SE, SI, SK, SM, TECHNOLOGY [US/US]; 77 Massachusetts Avenue, TR), OAPI (BF, BJ, CF, CG, CI, CM, GA, GN, GQ, GW, Cambridge, Massachusetts 02139 (US).
    [Show full text]